The Adam8 Knockout C2C12 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal cell population derived from the C2C12 myoblast line, featuring targeted disruption of the mouse Adam8 gene. This polyclonal knockout model enables loss-of-function studies without the clonal selection step, preserving population-level heterogeneity.
C2C12 cells are a well-characterized myoblast line originally isolated from the thigh muscle of a C3H mouse and represent a subclone of the C2 line. These cells serve as a skeletal muscle progenitor model capable of differentiating into multinucleated myotubes upon serum withdrawal, making them an essential system for investigating myogenesis, muscle differentiation, and regeneration. Their robust differentiation capacity and well-documented signaling responses render them particularly suited for examining the molecular mechanisms governing muscle development and disease.
ADAM8 (A Disintegrin and Metalloproteinase domain-containing protein 8) is an ectodomain sheddase that cleaves membrane-bound substrates including CD23 and L-selectin, releasing soluble ectodomains to modulate intercellular communication and adhesion. Its activity is stimulated by TNF-??, IL-1??, LPS, and phorbol esters, engaging NF-??B, ERK/MAPK, PI3K/Akt, and Notch signaling cascades that converge on integrin-mediated adhesion. By interacting with integrin ??v??3 and tetraspanin CD9, ADAM8 couples substrate shedding to actin cytoskeletal dynamics, promoting ERK phosphorylation and integrin activation to facilitate migration and inflammatory responses. In C2C12 myoblasts, knockout of ADAM8 abolishes the shedding of these substrates, thereby disrupting integrin-dependent adhesion and cytokine release, which impairs myogenic differentiation and reduces migratory competence.
In the C2C12 myoblast model, loss of ADAM8 function significantly alters the cellular program for muscle formation and repair. The absence of ADAM8-mediated shedding impairs the precise remodeling of cell?Ccell and cell?Cmatrix contacts required for myoblast alignment and fusion, leading to defective myotube formation. Additionally, disrupted integrin signaling and altered cytokine profiles compromise the ability of myoblasts to respond to regenerative cues and orchestrate inflammatory cross-talk within damaged muscle. This knockout model therefore provides a physiologically relevant platform to dissect how ADAM8 coordinates adhesive and proteolytic events essential for skeletal muscle homeostasis and regeneration.
Key research applications include investigating ADAM8’s role in myoblast fusion and differentiation, studying muscle regeneration mechanisms using injury or disease models, and exploring inflammatory myopathies where ADAM8-mediated shedding drives pathology. The polyclonal knockout cells are also valuable for drug target discovery in muscular dystrophy and cancer cachexia. Researchers can employ western blotting for shed substrates, transwell migration assays, myotube differentiation assays with MYH immunofluorescence, RT-qPCR for myogenic markers (MyoD, myogenin), flow cytometry for surface integrin levels, and gelatin zymography. For further details, please contact Ascent Research.
